Cystic fibrosis (CF) is the most common life shortening inherited disorder amongst Caucasians, and Pseudomonas aeruginosa pulmonary infections are the leading cause of mortality in CF patients. Treatment of patients infected with P. aeruginosa is complicated by the cell-to-cell signaling systems of this organism, which regulate biofilm formation, virulence genes, and antibiotic resistance genes such as efflux pumps. These mechanisms augment bacterial resistance to both antibiotics and host defense, causing a vicious cycle in which the body's immune system continuously mounts an unproductive assault on bacterial infection, resulting in chronic inflammation, tissue damage, and eventually respiratory failure. The co-founders of Agile Sciences, Inc. have discovered simple derivatives of sponge-derived marine natural products with unprecedented activity toward inhibiting and dispersing bacterial biofilms. These "Agilyte" compounds work synergistically with antibiotics to reduce bacterial growth and lower the MICs of antibiotics toward antibiotic-resistant bacteria. In preliminary work, Agilyte molecules have shown efficacy toward: 1) inhibiting and dispersing biofilms of P. aeruginosa at low-micromolar concentrations and 2) working synergistically with tobramycin to stop growth of P. aeruginosa in broth culture. The goal of this proposal is to assess the efficacy of Agile Sciences'lead Agilyte compounds in an in vivo mouse model of chronic P. aeruginosa infection developed by Dr. Richard Boucher at UNC Chapel Hill. To this end, the Specific Aims of this Phase I STTR Project are: 1. To perform advanced in vitro studies of 5 Agilyte compounds to inform lead compound selection for in vivo studies. 2. To evaluate efficacy of two lead compounds selected in Specific Aim #1 in the Boucher mouse model of chronic P. aeruginosa pulmonary infection. Dr. Laura Guogas, a microbiologist with expertise in cystic fibrosis pulmonary disease, will lead the in-house efforts t Agile Sciences as well as coordinate a team of expert collaborators. Testing of Agilyte molecules in vivo will be conducted in the laboratory of the co-PI, Dr. Richard Boucher, the Kenan Professor of Medicine and Cystic Fibrosis and Pulmonary Research and Treatment Center Director at UNC. Guidance on preclinical development of the Agilyte molecules will be provided by Dr. Ward Peterson, former Vice President of Research and Preclinical Development at Inspire Pharmaceuticals, a biotechnology company formerly focused on CF therapeutics. Agile co-founder Dr. Christian Melander will provide expertise on the biological properties of the Agilyte molecules. This multidisciplinary team will work cooperatively to assess the potential of Agile's novel technology to decrease P. aeruginosa proliferation under in vivo conditions relevant to the CF lung. If successfully developed, our proposed therapeutic has the potential to modulate the significant mortality and morbidity associated with CF disease through the eradication of chronic bacterial infection. PUBLIC HEALTH RELEVANCE: Developing therapeutics to treat cystic fibrosis (CF) is especially challenging due to the formation of communities of bacteria called biofilms in the lungs of cystic fibrosis patients. Agile Sciences is developing molecules that both inhibit and disperse bacterial biofilms and thus have the potential to significantly enhance the effectiveness of antibiotic therapy for CF. An improved treatment for CF has the potential to both extend the life expectancy and improve the quality of life for the hundreds of thousands of individuals worldwide who are living with CF.